DBT cells were plated
to be 80% confluent
at the start of the assay. Compounds were diluted as done for the
MHV assay and incubated with cells at 37 °C for 1 h. After 1
h, 50 μL of DMEM was added to the cells (T =
0); 45 min before harvest, lysis buffer was added to positive wells.
LDH activity in cell-free supernatants was measured at 10 h after
infection using the Sigma Tox7 kit as per the manufacturer’s
directions. A549-ACE2 cells were seeded at 20,000 cells per well 1
day prior to infection in 96-well plates. Cells were pretreated for
1 h and then mock-infected. Then, 2 h post-mock infection, the media
was removed, the monolayer was rinsed one time with PBS, and media
containing drug was added to each well. Typically,48 h after mock
infection, plates were centrifuged, and an aliquot of the cell culture
supernatant was removed. For LDH assays using Sigma Tox7 kit, the
clarified supernatant was transferred to a clean plate and assayed
following the manufacturer’s protocol.
to be 80% confluent
at the start of the assay. Compounds were diluted as done for the
MHV assay and incubated with cells at 37 °C for 1 h. After 1
h, 50 μL of DMEM was added to the cells (T =
0); 45 min before harvest, lysis buffer was added to positive wells.
LDH activity in cell-free supernatants was measured at 10 h after
infection using the Sigma Tox7 kit as per the manufacturer’s
directions. A549-ACE2 cells were seeded at 20,000 cells per well 1
day prior to infection in 96-well plates. Cells were pretreated for
1 h and then mock-infected. Then, 2 h post-mock infection, the media
was removed, the monolayer was rinsed one time with PBS, and media
containing drug was added to each well. Typically,48 h after mock
infection, plates were centrifuged, and an aliquot of the cell culture
supernatant was removed. For LDH assays using Sigma Tox7 kit, the
clarified supernatant was transferred to a clean plate and assayed
following the manufacturer’s protocol.